基于荧光微球免疫层析技术的河鲀毒素定量检测方法

摘要/Abstract

摘要： 建立了河鲀毒素的荧光微球免疫层析检测方法。基于抗体亲和位点保护标记方法制备了抗河鲀毒素抗体偶联荧光微球，方法简单稳定，所有结合分离步骤均在亲和柱中完成，不仅避免了亲和位点被破坏而且荧光信号更强，因此将检测灵敏度提高了8倍。所建立的河鲀毒素检测方法IC50为 18.4 ng/mL，线性范围为2 ng/mL -200ng/mL ( y = -0.15ln x +0.95，R2= 0.98)。检测河鲀样本的最低检出限为10μg/kg，平均相对标准偏差<25% (n=6)。该方法适用于热加工样品的检测，整个检测过程仅需15分钟，且无需使用大型设备，为河鲀餐前速测提供了有力的技术手段。

关键词: 河鲀毒素, 亲和位点保护, 荧光微球, 免疫层析

Abstract: A microsphere-based fluorescence immunochromatographic assay for detection of tetrodotoxin was established. A binding site protection procedure was developed for antibody labeling with microsphere-based fluorescence, which prevented damage to the antibody binding site and obtained stronger fluorescence signal. The combination and separation steps were efficiently completed in an affinity spin column. The sensitivity of the antibody-microsphere conjugate was eight times than it prepared by direct labeling. The Iimmunochromatographic assay exhibited dynamic linear range for detection of tetrodotoxin from 2 ng/mL to 100 ng/mL (y = -0.15ln x +0.95，R2= 0.98) with the IC50 of 18.4 ng/mL. The detection limit for tetrodotoxin was 10 μg/kg in puffer with the average RSD<25% (n=6). The assay method which did not need large-scale equipment could be applied in heat-processed products. The detection time for each sample was 15 min．

Key words: Tetrodotoxin, Binding site protection, fluorescent microspheres, immunochromatography

中图分类号:

TS207.3

张世伟王士峰姚添琪杨国武赖心田. 基于荧光微球免疫层析技术的河鲀毒素定量检测方法[J]. 食品科学, 0, (): 0-0.

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